Hydroxamic acid derivatives

ABSTRACT

Novel hydroxamic acid derivatives, e.g., of formula (I), wherein R 1 , R 2 , R 3  and R 4  are as defined, are found to be useful as pharmaceuticals, e.g., for the suppression of TNF release and the treatment of autoimmune and inflammatory diseases, e.g., multiple sclerosis and rheumatoid arthritis. Methods of making the compounds, novel intermediates, and pharmaceutical compositions comprising the compounds are provided.

This invention relates to novel hydroxamic acid derivatives and to theiruse as pharmaceuticals, e.g., in inhibiting matrix metalloproteinasessuch as collagenase, and in inhibiting TNF production, particularly fortreatment of diseases or conditions mediated by over-production of orover-responsiveness to TNFα.

Accordingly the invention provides compounds of Formula I

wherein

-   -   R₁ is lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl or        C₄-C₁₈aryl each of which is independently optionally substituted        by hydroxy, halogen, lower alkoxy, C₃-C₈cycloalkyl-lower alkoxy,        or C₄-C₁₈ aryl-lower alkoxy;    -   X is halogen, cyano, lower alkyl, halo-substituted lower alkyl,        C₄-C₁₈aryl, C₄-C₁₈aryl-lower alkyl, hydroxy, —OR₅, SR₅ or        —NR₆R₇, each of which is optionally substituted by halogen,        hydroxy, lower alkoxy, C₃-C₆cycloalkyl-lower alkoxy, or        C₄-C₁₈aryl-lower alkoxy    -   wherein        -   R₅ is hydrogen, lower alkyl, C₃-C₈cycloalkyl,            C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl and        -   R₆ and R₇ are independently H, lower alkyl, C₃-C₈cycloalkyl,            C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl;    -   Z is —CH₂—, —CHR₈—, —O—, —S—, or —N(R₈)—    -   wherein        -   R₈ is H, lower alkyl, C₃-C₈cycloalkyl,            C₃-C₁₈heterocycloalkyl, C₄-C₁₈aryl lower alkoxycarbonyl or            C₄-C₁₈aryloxycarbonyl, each of which is independently            optionally substituted by halogen, hydroxy, lower alkoxy,            C₃-C₆cycloalkyl-lower alkoxy, or C₄-C₁₈aryl-lower alkoxy;    -   A is hydrogen, —CR₁₀R₁₁-Q-R₁₂, —C(O)-Q-R₁₂ or —C(S)-Q-R₁₂    -   wherein        -   R₁₀ and R₁₁ are independently H, lower alky,            C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl each            of which is independently optionally substituted by halogen,            hydroxy, lower alkoxy, C₃-C₆cycloalkyl-lower alkoxy, or            C₄-C₁₈ aryl-lower alkoxy,        -   Q is —NR₈—, —S— or —O—, where R₈ is as defined above, and        -   R₁₂ is lower alkyl C₃-C₈cycloalkyl, C₄-C₁₈aryl,            C₄-C₁₈aryl-lower alkyl, each optionally substituted by            hydroxy, halogen, lower alkoxy, C₃-C₆cycloalkyl,            C₃-C₆cycloalkoxy, C₄-C₁₈aryl or C₄-C₁₈aryl-lower alkoxy; and    -   R₃ and R₄ are independently H or lower alkyl    -   n is 0 or 1,        and pharmaceutically-acceptable and -cleavable esters thereof        and acid addition salts thereof.

Preferably R₁ is H, lower alkyl or C₃-C₈cycloalkyl, each of which isoptionally substituted, preferably by hydroxy, halogen, lower alkoxy orC₄-C₈aryl-lower alkoxy.

R₁ as lower alkyl is preferably C₁-C₄lower-alkyl e.g. methyl, ethyl orpropyl, optionally substituted as defined above, for instance byphenyl-lower alkoxy e.g. as benzyloxymethyl.

R₁ as cycloalkyl is preferably C₃-C₆cycloalkyl, i.e. cyclopropyl,cyclobutyl, cyclopentyl or cyclohexyl, optionally substituted as definedabove.

X is preferably halogen, cyano, halo-substituted lower alkyl (e.g.trifluoromethyl), lower alkyl, or lower alkoxy, the latter two of whichare independently optionally substituted by halogen, hydroxy or loweralkoxy,

X as halogen may be fluorine, chlorine, bromine or iodine, and ispreferably fluorine or chlorine.

X as lower alkyl is preferably C₁-C₄ lower alkyl, e.g. methyl or ethyl,optionally substituted as defined above.

X as lower alkoxy is preferably C₁-C₄ lower alkoxy e.g. methoxy, orethoxy, optionally substituted as defined above.

Z is preferably —CH₂— or —N(R′₈)— wherein R′₈ is H, lower alkyl,C₄-C₈aryl (optionally substituted by halogen), lower alkoxycarbonyl orC₄-C₈aryloxycarbonyl.

Z as —N(R′₈)— is preferably

A is preferably H or —C(O)-Q′-R₁₂′ wherein

-   -   Q′ is S or O and R₁₂′ is lower alkyl, C₃-C₈cycloalkyl,        C₄-C₈aryl, each optionally substituted, by hydroxy, halogen,        lower alkoxy, C₃-C₆cycloalkyl, C₄-C₈aryl,    -   R₃ and R₄ are preferably H.    -   n is preferably 1.

Thus in a preferred embodiment the invention provides a compound offormula II

wherein

-   -   R¹′ is H, lower alkyl or C₃-C₈cycloalkyl, each of which is        optionally substituted by hydroxy, halogen, lower alkoxy or        C₄-C₈aryl-lower alkoxy;    -   X′ is halogen, cyano, lower alkyl, halo-substituted lower alkyl        or lower alkoxy, each of which is optionally substituted by        halogen, hydroxy or lower alkoxy;    -   Z′ is —CH₂— or —N(R′₈)— wherein R′₈ is H, lower alkyl, C₄-C₈aryl        (optionally substituted by halogen), lower alkoxycarbonyl or        C₄-C₈aryloxycarbonyl;    -   A′ is H or —C(O)-Q′-R₁₂′ wherein Q′ is —S— or —O— and R₁₂′ is        lower alkyl, C₃-C₈ cycloalkyl, C₄-C₈aryl, each optionally        substituted by hydroxy, halogen, lower alkoxy, C₃-C₈cycloalkyl,        or C₄-C₈aryl, and        pharmaceutically acceptable and cleavable esters thereof and        acid addition salts thereof.

In particularly preferred compounds of Formula II the substituents R₁′,X′, Z′, Q′ and R₁₂′ have the following meanings, independently or in anylogical combination thereof:

-   -   R₁′ is ethyl, hydroxymethyl or benzyloxymethyl.    -   X′ is halogen, e.g. chlorine, or lower alkoxy e.g. methoxy.    -   Z′ is —O—,

-   -    —CH₂—, or N-BOC.    -   A′ is H or —C(O)-Q″-R″₁₂    -   wherein    -   Q″ is O, and    -   R″₁₂ is methyl, ethyl, propyl, isopropyl, cyclopropyl,        iso-butyl, cyclopropylmethyl, sec. butyl, 1,2-dimethylpropyl,        3-methylbutyl, 2-methylbutyl, 2-methoxy-ethyl, 3-methoxy-propyl,        3-isopropoxy-propyl, 2-methoxy-1-methyl-ethyl,        4-hydroxy-cyclohexyl, 2-hydroxy-1-methyl-2-phenyl-ethyl, benzyl,        4-fluorophenyl, 2-hydroxypropyl, 2-methoxy-1-methyl-ethyl,        4-hydroxycyclohexyl, 1-phenylethyl, phenyl, phenylethyl, and        2-hydroxy-propyl.

Pharmaceutically acceptable salts of the acidic compounds of theinvention are salts formed with bases, namely cationic salts such asalkali and alkaline earth metal salts, such as sodium, lithium,potassium, calcium, magnesium, as well as ammonium salts, such asammonium, trimethyl-ammonium, diethylammonium, andtris-(hydroxymethyl)-methyl-ammonium salts.

Similarly acid addition salts, such as of mineral acids, organiccarboxylic and organic sulfonic acids e.g. hydrochloric acid,methanesulfonic acid, maleic acid, are also possible provided a basicgroup, such as pyridyl, constitutes part of the structure.

Pharmaceutically acceptable esters are, for instance, ester derivativeswhich are convertible by solvolysis or under physiological conditions tothe free carboxylic acids of formula I. Such esters are e.g. lower alkylesters (such as the methyl or ethyl ester), carboxy-lower all esterssuch as the carboxymethyl ester, nitrooxy-lower alkyl esters (such asthe 4-nitrooxybutyl ester), and the like.

The compounds of formulae I and II, depending on the nature ofsubstituents, may possess one or more asymmetric carbon atoms. Theresulting diastereomers and enantiomers are encompassed by the instantinvention. Preferably, however, e.g. for pharmaceutical use inaccordance with the invention, the compounds of formulae I and II areprovided in pure or substantially pure epimeric form, e.g. ascompositions in which the compounds are present in a form comprising atleast 90%, e.g. preferably at least 95% of a single epimer (i.e.comprising less than 10%, e.g. preferably less than 5% of other epimericforms).

Preferred are the compounds of formula I′ and formula I″

wherein the symbols are as defined above.

Above and elsewhere in the present description the following terms havethe meanings given below:

The term “lower” referred to above and hereinafter in connection withorganic radicals or compounds respectively defines a compound or radicalwhich may be branched or unbranched with up to and including 7,preferably up to and including 4 and (as unbranched) one or two carbonatoms.

A lower alkyl group is branched or unbranched and contains 1 to 7 carbonatoms, preferably 1-4 carbon atoms. Lower alkyl represents, for example,methyl, ethyl, propyl, butyl, isopropyl or isobutyl.

A lower alkoxy (or alkyloxy) group preferably contains 1-7 carbon atoms,advantageously 1-6 carbon atoms, and represents for example ethoxy,propoxy, isopropoxy, isobutoxy, preferably methoxy. Lower alkoxyincludes cycloalkyloxy and cyloalkyl-lower alkyloxy.

Halogen (halo) preferably represents chloro or fluoro but may also bebromo or iodo.

Aryl represents carbocyclic or heterocyclic aryl.

Carbocyclic aryl represents monocyclic, bicyclic or tricyclic aryl, forexample phenyl or phenyl mono-, di- or tri-substituted by one, two orthree radicals as hereinbefore defined.

Preferred as carbocyclic aryl is naphthyl, phenyl or phenyl mono- ordisubstituted as hereinbefore defined.

Heterocyclic aryl represents monocyclic or bicyclic heteroaryl, forexample pyridyl, indolyl, quinoxalinyl, quinolinyl, isoquinolinyl,benzothienyl, benzofuranyl, benzopyranyl, benzothiopyranyl, furanyl,pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl,pyrazolyl, imidazolyl, thienyl, or any said radical substituted,especially mono- or di-substituted, as hereinbefore defined.

Cycloalkyl represents a saturated cyclic hydrocarbon optionallysubstituted by lower alkyl which contains 3 to 8 ring carbons and isadvantageously cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyloptionally substituted as hereinbefore defined.

Amino may be optionally substituted, e.g. by lower alkyl.

Heterocyclyl represents a saturated cyclic hydrocarbon containing one ormore, preferably 1 or 2, hetero atoms selected from O, N or S, andpreferably from 3 to 10, more preferably 5 to 8, ring atoms; forexample, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyrrolyl,piperidinyl, piperazinyl or morpholino; all of which may be optionallysubstituted, for instance as hereinbefore defined.

Aryl-lower alkyl represents preferably (carbocyclic aryl or heterocylicaryl)-lower alkyl.

Carbocyclic aryl-lower alkyl preferably represents aryl-straight chainor -branched C₁₋₄-alkyl in which carbocyclic aryl has meaning as definedabove, e.g. benzyl or phenyl-(ethyl, propyl or butyl), eachunsubstituted or substituted preferably on the phenyl ring ashereinbefore defined for carbocyclic aryl above, advantageouslyoptionally substituted benzyl.

Heterocyclic aryl-lower alkyl represents preferably straight chain orbranched heterocyclic aryl-C₁₋₄-alkyl in which heterocyclic aryl hasmeaning as defined above, e.g. 2-, 3- or 4-pyridylmethyl or (2, 3- or4-pyridyl)-(ethyl, propyl or butyl); or 2- or 3-thienylmethyl or (2- or3-thienyl)-(ethyl, propyl or butyl); 2-, 3- or 4-quinolinylmethyl or(2-, 3- or 4-quinolinyl)-(ethyl, propyl or butyl); or 2- or4-thiazolylmethyl or (2- or 4-thiazolyl)-(ethyl, propyl or butyl).

Cycloalkyl-lower alkyl represents e.g. (cyclopropyl- orcyclobutyl)-(methyl or ethyl).

The invention includes the following compounds:

-   3(S)-(4-Chloro-phenyl)-2(S)-ethyl-N-hydroxymorpholin-4-yl-4-oxo-butyrmide;-   2(R)-Benzyloxymethyl-4-[4-(4chloro-phenyl)-piperazin-1-yl]-N-hydroxy-3(S)-(4-methoxy-phenyl)-4-oxo-butyramide;-   2(R)-Benzyloxymethyl-N-hydroxy-3(S)-(4-methoxy-phenyl)-4-oxo-4-piperidin-1-yl-butyramide,-   N-Hydroxy-2(R)-hydroxymethyl-3(S)-(4-methoxy-phenyl)-4-oxo-piperidin-1-yl-butyramide;-   (S)4-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-3-isobutylcarbamoyl-piperazine-1-carboxylic    acid .tert.-butyl ester;-   (S)-1-[(2S,3S-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperazine-2-carboxylic    acid isobutyl-amide trifluoro-acetate;-   1-[4-Benzyloxy-3(R)-hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-butyryl]-piperidine-2(S)-carboxylic    acid methylamide;-   1-[4-Hydroxy-3(R)-hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-butyryl]-piperidine-2(S)-carboxylic    acid methylamide;-   1-[3(S)-Hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-pentanoyl]-piperidine-2(S)-carboxylic    acid methylamide;-   (S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylic    acid cyclopropylamide;-   (S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylic    acid (2-methoxy-ethyl)-amide;-   (S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylic    acid (4-hydroxy-cyclohexyl)-amide;-   (S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylic    acid benzylamide;-   (S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylic    acid (4-fluoro-phenyl)-amide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylic    acid isopropylamide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbarnoyl-pentanoyl]-piperidine-2-carboxylic    acid cyclopropylamide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylic    acid (3-isopropoxy-propyl)-amide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylic    acid (4-hydroxy-cyclohexyl)-amide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylc    acid benzylamide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylic    acid phenylamide;-   1-[3(S)-Hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-pentanoyl]-pyrrolidine-2(S)-carboxylic    acid phenylamide;-   (S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-pyrroldine-2-carboxylic    acid ((S)-2-hydroxy-propyl)-amide.

The compounds of formula I, II, I′ and I″ and the specific compoundslisted above are hereinafter referred to as Compounds of the Invention.

Compounds of the Invention of Formula I are obtained by conversion of acorresponding free carboxylic acid derivative of formula V

wherein the symbols are as defined above, to the correspondinghydroxamic acid derivative of formula I.

An acid of formula V is converted to an activated acid. For example anacid of formula V is dissolved in an inert solvent, such asdichloromethane, HOPO and 4-(2-isocyano-ethyl)-morpholine are addedsuccessively. The activated acid is then treated with a protected aminoalcohol; for example, a silyl protected amino alcohol e.g. TMSONH₂, toyield a compound of formula I

The free carboxylic acid derivatives of formula V are prepared byoxidation of a corresponding olefin derivative of formula VI

wherein the symbols are as defined above.For example the crude olefin of formula VI is dissolved in a polar inertnon-aqueous solvent such as lower aliphatic alcohol, advantageouslymethanol and preferably cooled e.g. −85° C.-−65° C. Ozone is introducedpreferably at a reduced temperature e.g. −80° C.-−60° C. The solution isthen purged with an inert gas such as nitrogen and treated withdimethylsulfide. The solvent is removed and the crude oil redissolved ina polar inert solvent such as lower aliphatic alcohol, advantageouslyt-butanol. A solution of sodium dihydrogenphosphate monohydrate in apolar solvent such as water is added, followed by 2-methyl-2-butene andsodium chlorite in a polar solvent such as water. When the reaction hasceased the reaction mixture is treated with sodium sulfite in a polarsolvent such as water to yield a compound of the formula V

The olefin derivatives of formula VI are advantageously prepared bycoupling of olefin derivatives of formula VII

with a N-heterocycle derivative of formula VIII

wherein the symbols are as defined above.For example a N-heterocycle derivative of formula VIII and an olefinderivative of formula VII are dissolved in a polar solvent such as DMPand preferably cooled e.g. −10° C.-10° C. HOBT, lower alkylamine such astriethylamine and a coupling reagent such as EDC are added successively.The reaction mixture is allowed to warm to room temperature. To yield acompound of formula VI

The olefin precursors of formula VII are prepared by rearrangement of aprecursor compound of formula IX

wherein the symbols are as defined aboveFor example hexamethyl disilazan is dissolved under an inert atmospheresuch as argon in a polar inert non-aqueous solvent such as THF andpreferably cooled e.g. −10° C.-10° C. A lower alkyllithium such asn-Butyl lithium in an inert solvent such as hexane is added. Thereaction mixture is preferably cooled further e.g. −68° C.-−88° C. and alower alkylchlorosilane such as trimethyl chlorosilane is added,followed by addition of a solution of a compound of formula IX in apolar inert non-aqueous solvent such as THF. Titanium tetrachloride inand inert solvent such as dichloromethane is added and the reactionmixture preferably allowed to warm up e.g. to room temperature to yielda compound of formula VII

The derivatives of formula VIII are prepared by a deprotection of aprotected carboxylic acid of formula XII

wherein the symbols are as defined above or in the case of {circlearound (P)} as defined below. {circle around (P)} is a carboxylic acidprotecting group which may be a silyl protecting group (e.g.tertiarybutyldimethylsilyl or trimethylsilyl), or any other carboxylicacid protecting group commonly known in the art.For example the amide of formula XII is treated with a lower alkanoicacid such as trihalo acetic acid for example trifluro-acetic acid TFA ata reduced temperature preferably −10° C.-10° C. The mixture isevaporated and HCl is added to yield a compound of formula XII

Compounds of the Invention, as defined above, e.g., of formula I, II, I′and I″ and particularly as exemplified, in free or pharmaceuticallyacceptable ester and salt form, exhibit pharmacological activity and areuseful as pharmaceuticals, e.g. for therapy, in the treatment ofdiseases and conditions as hereinafter set forth.

As discussed in the test procedures described below, the Compounds ofthe Invention are potent inhibitors of TNFα release, are orally active,and are not cytotoxic at their effective doses. The Compounds of theInvention also inhibit collagenase and stromelysin at concentrations offrom 0.3 to 10 nM and are shown to be good inhibitors of mechanicalhyperalgesia. The Compounds of the Invention tested further show oralactivity in vivo at dosages of less than 10 mg/kg in LPS induced TNFαrelease in the rat, and appear to be well tolerated at such dosages.

Test Procedure 1: Inhibition of TNF release

Mononuclear cells are prepared from the peripheral blood of healthyvolunteers using ficoll-hypaque density separation according to themethod of Hansell et al., J. Imm. Methods (1991) 145: 105, and used at aconcentration of 10⁵ cells/well in RPMI 1640 plus 10% FCS. Cells areincubated with serial dilutions of the test compounds for 30 minutes at37° C. prior to the addition of IFNγ (100 U/ml) and LPS (5 μg/ml) andsubsequently further incubated for three hours. Incubation is terminatedby centrifugation at 1400 RPM for 10 min. TNFα in the supernatant ismeasured using a commercial ELISA (Innotest hTNFα, available fromInnogenetics N.V., Zwijnaarde, Belgium). Compounds of the Invention aretested at concentrations of from 0 to 10 μM. Exemplified compounds offormula I, especially of formula Ia, suppress TNF release in this assaywith an IC₅₀ of from about 50 nM to about 5 μM.

Test Procedure 2: Cytotoxicity

Cytotoxicity is determined on THP1 cells (5×10⁴/well) which areincubated in the presence of IFNγ (100 U/ml) and LPS (5 μg/ml) andpresence and absence of test compound for 24 hours at 37° C. Percentagesof living and dead cells are assessed by a colorimetric readout (MTT),which measures mitochondrial dehydrogenase enzymes in living cells, asdescribed in Mosman, J. Imm. Methods (1983) 65: 55. Compounds of theInvention tested show less than 50% cytotoxicity at a concentration of10 μM, showing that the Compounds of the Invention are not cytotoxic atconcentrations sufficient to suppress TNF.

Test Procedure 3: Collagenase Inhibition

Collagenase inhibition is determined using active collagenase with thethiopeptide MMP-substrate described in Stein and Izquierdo-Martin, Arch.Biochem. Biophys. 308 (1994) pp. 274-277. Test compound is incubatedwith the collagenase prior to the addition of the substrate at pH 6.5,25° C. in 2-morpholinoethanesulphonic acid (50 mM) buffer with 10 mMCaCl₂. The absorbance is recorded at 405nm at regular intervals for aperiod of 40 minutes. The inhibitory activity of the test compound isdetermined as a function of the collagenase activity in the control inthe presence and absence of the test compound. Compounds of theInvention show significant dose dependent inhibition of collagenase atlow nM concentrations, e.g., below 10 nM.

Test Procedure 4: Oral Bioavailability

The assay of the preceding example is standardized by measuring activityof varying known concentrations of a particular test compound and usedto measure the concentration of test compound in plasma following oraladministration. Test compounds are administered orally to conscious ratsat a dosage of 10 mg/kg. Blood samples are taken from the cut tip of thetail at 30, 60, 120, and 240 minutes from oral administration. Theplasma is subjected to trichloroacetic acid extraction. The extract istested in the above collagenase inhibition assay to obtain an estimateof the concentration of drug present in the plasma Compounds of theInvention show good oral bioavailability, with plasma concentrations of300-5000 nM after 30 minutes and 50-500 nM after 240 minutes. Thus,pharmaceutically effective plasma levels (as shown in Test Example 1 and3) are readily achievable with oral administration at manageabledosages, e.g., 10 mg/kg. Moreover, the plasma levels obtained are wellbelow the cytotoxic level, and the rats were not observed to show anyadverse effects at this dosage.

Test Procedure 5: Effect on Mechanical Hyperalgesia and Allodynia

The effect on mechanical hyperalgesia and allodynia is determined usinga model of neuropathic pain in rat (Seltzer et al, 1990). A 7-0 silksuture is inserted into the sciatic nerve of one of the hind-paws,usually the left, causing mechanical hyperalgesia and allodynia to beinduced. Mechanical hyperalgesia is measured by the paw withdrawalthresholds of the hind-paws to increasing pressure stimulus using anAnalgesymeter. Mechanical allodynia is measured by the withdrawalthresholds of the hind-paws to non-noxious mechanical stimuli appliedusing von Frey hairs. Test compound or vehicle (20% cremaphor/water) wasadministered orally twice daily for 7 days commencing from the day ofsurgery. Paw withdrawal thresholds of vehicle treated animals wasapproximately 60 g and paw withdrawal thresholds of test compoundtreated animals increased, with repeated dosing, to 80-90 g. Three daysafter the end of test compound administration the paw withdrawalthreshold of the test compound treated animals remained elevated abovethat of the vehicle treated animals. Treatment with test compound priorto surgery and a further single treatment with the test compound aftersurgery had a significant degree of inhibition of hyperalgesia lastingseveral days. Compounds of the invention tested were also effective inthe inhibition of established mechanical hyperalgesia. After just fourdays of administration of test compound, to animals with establishedmechanical hyperalgesia, there was a significant increase in the pawwithdrawal threshold compared to the vehicle treated animals.

Accordingly, Compounds of the Invention have pharmaceutical utility asfollows:

Compounds of the Invention are useful for the prophylaxis and treatmentof diseases or pathological conditions mediated by TNF, especially TNFα,e.g., inflammatory conditions, autoimmune diseases, severe infections,and organ or tissue transplant rejection, e.g. for the treatment ofrecipients of heart, lung, combined heart-lung, liver, kidney,pancreatic, skin or corneal transplants and for the prevention ofgraft-versus-host disease, such as following bone marrow transplants.

Compounds of the Invention are particularly useful for the treatment,prevention, or amelioration of autoimmune disease and of inflammatoryconditions, in particular inflammatory conditions with an aetiologyincluding an autoimmune component such as arthritis (for examplerheumatoid arthritis, arthritis chronica progrediente and arthritisdeformans) and rheumatic diseases. Specific auto-immune diseases forwhich Compounds of the Invention may be employed include autoimmunehaematological disorders (including e.g. hemolytic anaemia, aplasticanaemia, pure red cell anaemia and idiopathic thrombocytopenia),systemic lupus erythematosus, polychondritis, sclerodoma, Wegenergranulamatosis, dermatomyositis, chronic active hepatitis, myastheniagravis, psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmuneinflammatory bowel disease (including e.g. ulcerative colitis andCrohn's disease), endocrine ophthalmopathy, Graves disease, sarcoidosis,multiple sclerosis, primary biliary cirrhosis, juvenile diabetes(diabetes mellitus type I), uveitis (anterior and posterior),keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitiallung fibrosis, psoriatic arthritis and glomerulonephritis (with andwithout nephrotic syndrome, e.g. including idiopathic nephrotic syndromeor minimal change nephropathy).

Compounds of the Invention are also useful for the treatment,prevention, or amelioration of asthma, bronchitis, pneumoconiosis,pulmonary emphysema, and other obstructive or inflammatory diseases ofthe airways.

Compounds of the Invention are useful for treating undesirable acute andhyperacute inflammatory reactions which are mediated by TNF, especiallyby TNFα, e.g., acute infections, for example septic shock (e.g.,endotoxic shock and adult respiratory distress syndrome), meningitis,pneumonia; and severe burns; and for the treatment of cachexia orwasting syndrome associated with morbid TNF release, consequent toinfection, cancer, or organ dysfunction, especially AIDS-relatedcachexia, e.g., associated with or consequential to HIV infection.

In addition to inhibiting the release of TNF, especially TNFα throughthe suppression of TNF convertase, Compounds of the Invention are alsoinhibitors of matrix metalloproteinases, e.g., collagenase, stromelysinand gelatinases, and hence useful for the indications known forcollagenase inhibitors or other matrix metalloproteinase inhibitors,e.g., treatment of various pathological conditions of the skin, bones,and connective tissues, e.g., rheumatoid arthritis, psoriasis, psoriaticarthritis, osteoporosis, osteoarthritis, periodontitis, gingivitis, andcorneal ulceration; for the treatment of cardiovascular disease, e.g.,atherosclerosis, and coronary angioplasty; for the prevention of tumorcell metastasis and invasion and in inducing fibrosis of tumors, e.g.,in the treatment of cancer; and for the prevention of neurodegenerativedisorders, e.g., Alzheimer's disease.

Compounds of the Invention in addition to inhibiting the release of THF,especially TNFα through the suppression of TNF convertase, and theinhibition of matrix metalloprotinases, are also inhibitors ofneuropathic pain and associated hyperalgesia and are hence useful forthe indications known for neuropathic pain inhibitors, e.g., treatmentof neuropathic pain and associated hyperalgesia, including trigeminaland herpetic neuralgia, diabetic neuropathic pain, migraine, causalgiaand defferentation syndromes such as brachial plexus avulsion.

For the indications described above the appropriate dosage will, ofcourse, vary depending, for example, on the particular Compound of theInvention employed, the subject to be treated, the mode ofadministration and the nature and severity of the condition beingtreated. However, in general, satisfactory results in animals areobtained at daily dosages of from about 0.1 to about 100 mg/kg/day p.o.In larger mammals, for example humans, an indicated daily dosage is inthe range of from about 5 to about 1000 mg of Compound of the Inventionadministered orally once or, more suitably, in divided dosages two tofour times/day.

Compounds of the Invention may be administered by any conventionalroute, e.g. orally, for example in the form of solutions for drinking,tablets or capsules or parenterally, for example in the form ofinjectable solutions or suspensions. Normally for systemicadministration oral dosage forms are preferred, although for someindications Compounds of the Invention may also be administeredtopically or dermally, e.g. in the form of a dermal cream or gel or likepreparation or, for the purposes of application to the eye, in the formof an ocular cream, gel or eye-drop preparation; or may be administeredby inhalation, e.g., for treating asthma. Suitable unit dosage forms fororal administration comprise e.g. from 25 to 250 mg Compound of theInvention per unit dosage.

In accordance with the foregoing the present invention also provides ina further series of embodiments:

-   -   A. A method of inhibiting production of soluble TNF, especially        TNFα, or of reducing inflammation in a subject (i.e., a mammal,        especially a human) in need of such treatment which method        comprises administering to said subject an effective amount of a        Compound of the Invention, or a method of treating any of the        above mentioned conditions, particularly a method of treating an        inflammatory or autoimmune disease or condition, e.g., multiple        sclerosis or rheumatoid arthritis, or alleviating one or more        symptoms of any of the above mentioned conditions.    -   B. A method of inhibiting neuropathic pain and associated        hyperalgesia in a subject (i.e. a mammal, especially a human) in        need of such treatment which method comprises administering to        said subject an effective amount of a compound of the invention,        or a method of treating any of the above mentioned disease or        conditions, e.g., neuropathic pain and associated hyperalgesia,        diabetic neuropathic pain or migraine, or alleviating one or        more symptoms of any of the above mentioned conditions.    -   C. B. A Compound of the Invention for use as a pharmaceutical,        e.g. for use as an immunosuppressant, antiinflammatory or        neuropathic pain relief agent or for use in the prevention,        amelioration or treatment of any disease or condition as        described above, e.g., an autoimmune, inflammatory or        neuropathic pain disease or condition.    -   D. C. A pharmaceutical composition comprising a Compound of the        Invention in association with a pharmaceutically acceptable        diluent or carrier, e.g., for use as an immunosuppressant,        anti-inflammatory or neuropathic pain relief agent or for use in        the prevention, amelioration or treatment of any disease or        condition as described above, e.g., an autoimmune, inflammatory        or neuropathic pain disease or condition.    -   E. D. Use of a Compound of the Invention in the manufacture of a        medicament for use as an immunosuppressant, anti-inflammatory or        neuropathic pain relief agent or for use in the prevention,        amelioration or treatment of any disease or condition as        described above, e.g., an autoimmune, inflammatory or        neuropathic pain disease or condition.        Abbreviations used herein in relation to the invention:

-   BOC: Benzyloxycarbonyl

-   DCC: Dicyclohexyl-carbodiimide

-   DMAP: Dimethyl-pyridin-4-yl-amine

-   DMF: N,N-Dimethyl formamide

-   EDC: (3-Dimethylamino-propyl)-ethyl-carbodiimide hydrochloride

-   HCl: Hydrochloric acid

-   HOBT: Benzotriazol-1-ol

-   HOPO: 1-Oxy-pyridin-2-ol

-   MEI: 4-(2-Isocyano-ethyl)-morpholine

-   NaOH: Sodium hydroxide

-   TBME: t-Butyl methyl ether

-   TFA: Trifluoro-acetic acid

-   THF: Tetrahydrofuran

-   TMSONH2: O-(Trimethylsilyl)-hydroxylamine

Synthesis of the Intermediates VII′

The intermediates of formula VII′ were prepared by the general Method IAwhich comprises the coupling of a suitable allylic alcohol with thecorresponding phenyl-acetic acid, followed by an adapted Ireland-Claisenrearrangement and optical resolution of the enantiomers using(S)-(−)-1-phenyl-ethylamine.

Method IA is illustrated by the following representative examples.

EXAMPLE I1 3(R)-Benzyloxymethyl-2(S)-(4-methoxy-phenyl)-pent-4-enoicacid (I1)

Step A: Prop-2-ynyloxymethyl-benzene (I1a)

Tetrabutylammonium bromide (19.34 g, 600 mmol) was added to a solutionof benzylbromide (71.3 ml, 600 mmol) and propargyl alcohol (35.5 ml, 600mmol) in 270 ml of toluene and the mixture was heated to 50° C. Asolution of sodium hydroxide (24 g, 600 mmol) in 55 ml of water was thenadded dropwise over a period of 1 hour and stirring was continued foranother 3 hours. The reaction mixture was then cooled to 25° C. andtransferred into a separatory funnel. The aqueous layer was removed andthe organic layer was washed three times with 150 ml of brine. Residualwater was removed by repeated azeotropic distillation using toluene. Apale yellow liquid was obtained. Yield: 81.9 g (93.4 %).

MS (EI): 145 [M−H]⁻ ¹H NMR (CDCl₃), δ (ppm): 7.29-7.40 (m, 5H), 4.64 (s,2H), 4.20 (d, 2H), 2.49 (t, 1H).

Step B: 4-Benzyloxy-but-2-yn-1-ol (I1b)

The crude alkyne I1 (81.9 g, 560 mmol) was dissolved in 500 ml of THFand cooled to −78° C. A 2.5M solution of n-hexyl lithium (250 ml, 625mmol) was added dropwise over a period of 2 hours, so that thetemperature did not exceed −70° C. Stirring was continued for 3 hours at−78° C. and then solid paraformaldehyde (20.18 g, 672 mmol) was added.The dry ice-bath was removed and the reaction allowed to warm up to roomtemperature over night (approx. 20 hours). The mixture was diluted with600 ml of TBME and extracted with 400 ml of an ammonium chloridesolution (20%). The aqueous layer was extracted once more with 200 ml ofTBME and the combined organic fractions were washed three times with 400ml of a brine, dried over anhydrous sodium sulfate and evaporated togive an orange liquid. Yield: 87.9 g (89%).

¹H NMR (CDCl₃), δ (ppm): 7.25-7.40 (m, 5H), 4.62 (s, 2H), 4.35 (t, 2H),4.24 (t, 2H).

Step C: trans-4-Benzyloxy-but-2-en-1-ol (I1c)

Commercial Red-Al (70% in toluene) (85.2 g, 295 mmol) was added dropwiseto a cooled (−2° C.) solution of crude alcohol I1b (40 g, 227 mmol)dissolved in 200 ml of THF. The temperature was kept below 5° C. duringthe addition. After 1 hour the reaction had ceased and a fine suspensionformed which was added dropwise to 300 ml of ice cold 20% sulfuric acid.The resulting suspension was diluted with 250 ml of toluene and stirredvigorously for 30 minutes. The two layers were then separated and theaqueous phase extracted again with of toluene. The combined organiclayers were washed with saturated sodium bicarbonate solution and brine,evaporated and dried by repeated azeotropic distillation using toluene.An orange brown liquid was obtained which was distilled under reducedpressure (110° C., 0.05 mbar). Yield: 32.65 g (80.7%).

¹H NMR (CDCl₃), δ (ppm): 7.15-7.35 (m, 5H), 5.65-5.90 (m, 2H), 4.46 (s,2H), 4.09 (br. d, 2H), 3.97 (d, 2H).

Step D: trans-(4-Methoxy-phenyl)-acetic acid 4-benzyloxy-but-2-enylester (I1d)

DMAP (3.425 g, 28 mmol) was added to a suspension of the alcohol I1c(100 g, 560 mmol) and (4-methoxy-phenyl)-acetic acid (93.05 g, 560 mmol)in 425 ml of toluene. After 15 minutes of stirring a yellow solution wasformed. A solution of DCC (115.55 g, 560 mmol) in 225 ml of DMP wasadded dropwise during 25 minutes. Eventual cooling was required to keepthe temperature between 23 and 27° C. Stirring was continued for 1 hour.Then, heptane (225 ml) was added, the suspension was cooled to −5° C.,filtered and washed with 300 ml of cold heptane. The filtrate was washedtwice with 1000 ml of water, filtered through basic aluminium oxide(ALOX, 500 g) and evaporated to give a yellow oil. Yield: 161.4 g (88%).

MS (EI): 349 [M+Na]⁺ ¹H NMR (CDCl₃), δ (ppm): 6.60-7.20 (m, 9H, 5.65 (m,2H), 4.40 (t, 2H), 4.31 (s, 2H), 3.82 (d, 2H), 3.58 (s, 3H), 3.38 (s,2H).

Step E: 3-Benzyloxymethyl-2-(4-methoxy-phenyl)-pent-4-enoic acid (I1e)

A solution of hexamethyl disilazane (38.4 ml, 184.3 mmol) in 100 ml ofTHF is cooled to 0° C. under nitrogen. Hexamethyl lithium (26.6% inhexane, 63.65 g, 184 mmol) was added over 15 minutes at a temperaturebelow 4° C. The mixture was cooled to −78° C. and trimethyl chlorosilane(22.65 ml, 184 mmol) was added dropwise over 15 minutes followed by asolution of the ester I1d (50 g, 153.2 mmol) in 50 ml of THF (over 45minutes). Finally, titanium tetrachloride (1M in toluene, 310 μl, 0.31mmol) was injected and the brown mixture was allowed to warm to 20° C.over a period of 90 minutes. Stirring was continued for another hour andthe resulting mixture was poured into 400 ml of 1N-NaOH. The aqueouslayer was separated and the organic layer was extracted with another 70ml portion of 1N-NaOH. The combined aqueous layers were acidified with390 ml of 5%-HCl and extracted with toluene (300 ml) which was thenevaporated to give the title compound as a racemic mixture ofdiastereomers (syn/anti ratio of 13/1). Yield: 38.35 g (76.7%)

Step F: 3(R)-Benzyloxymethyl-2(S)-(4-methoxy-phenyl)-pent-4-enoic acid(I1)

A solution of crude acid I1e (45.17 g, 138.3 mmol) in 1850 ml of ethylacetate was treated with of (S)-(−)-1-phenyl-ethylamine (18.4 g, 152.2mmol). A white precipitate was formed immediately, which dissolved uponheating to reflux. The solution was allowed to cool down to 20° C. overa period of 4 hours. The formed crystals (25.35 g, 41%) were filteredoff, dried under vacuum and recrystallized from 1125 ml of ethyl acetateto give 15.7 g (25%) of enantiomerically pure salt. This material wassuspended in 350 ml of toluene and extracted with 100 ml of 1N-HCl. Theorganic layer was washed twice with water and evaporated to give aviscous oil which solidified upon standing. Yield: 11.3 g (99%).

MS (EI): 325 [M−H]⁻ ¹H NMR (CDCl₃), δ (ppm): 6.70-7.40 (m, 9H), 5.40 (m,1H), 4.87 (m, 2H), 4.43 (s, 2H), 3.71 (s, 3H, overlapping with d, 1H),3.50 (d×d, 1H), 3.37 (d×d, 1H), 3.09 (m, 1H).

EXAMPLE I2 2(S)-(4-Chloro-phenyl)-3(R)-ethyl-pent-4-enoic acid (I2)

Step A: (4-Chloro-phenyl)-acetic acid pent-2-enyl ester (I2a)

Trans-2-penten-1-ol (49.6 ml; 487.6 mmol) and (4-chloro-phenyl)-aceticacid (83.2 g; 487.6 mmol) were dissolved in 1200 ml of dichloromethane.After addition of EDC (140.2 g, 731 mmol) and DMAP (11.9 g, 97.5 mmol),the reaction mixture was stirred for 18 hours at room temperature. Afterevaporation under reduced pressure, the residue was dissolved withdiethyl ether, washed with water, brine and dried over anhydrous sodiumsulfate. Evaporation gave 136 g of an yellow oil, which was furtherpurified by flash-chromatography (silica gel; hexane/ethyl acetate 9:1).Yield: 68 g (59%).

¹H-NMR (CDCl₃): δ (ppm) 7.25 (d, 2H), 7.15 (d, 2H), 5.68-5.78 (m, 1H),5.42-5.52 (m, 1H), 4.47 (d, 2H), 3.52 (s, 2H), 2.0 (q, 2H), 0.92 (t,3H).

Step B: 2-(4-Chloro-phenyl)-3-ethyl-pent-4-enoic acid (I2b)

Hexamethyl disilazan (20.3 g, 126 mmol) was dissolved under argon in 200ml of dry THF and cooled to 0° C. n-Butyl lithium (1.6M in hexane, 79ml, 126 mmol) was added dropwise and stirred for 10 minutes at 0° C. Thereaction mixture was cooled to −78° C. and trimethyl chlorosilane (16ml, 126 mmol) was added via syringe, followed by addition of a solutionof I2a (20 g, 83.8 mmol) in 50 ml of THF. After stirring for 1 hour at−78° C., titanium tetrachloride (1M in dichloromethane, 1.7 ml, 1.7mmol) was added via syringe and the reaction mixture was allowed to warmup to room temperature, and was stirred for 1 hour. The mixture wasevaporated and the residue was dissolved in diethyl ether and extractedtwice with 1N-NaOH. The aqueous layer was acidified with 1N-HCl andextracted with diethyl ether. The organic layers were dried overanhydrous sodium sulfate and evaporated under reduced pressure. Yield:17.3 g of white crystals (87%) as a racemic mixture of diastereomers(syn/anti ratio of 9/1).

Step C: 2(S)-(4-Chloro-phenyl)-3(R)-ethyl-pent-4-enoic acid (I2)

A hot solution of the racemate I2b (17.3 g, 72.5 mmol) in ethanol (250ml) was treated with (S)-(−)-1-phenyl-ethylamine (10.1 ml, 72.5 mmol).After slowly cooling down, the precipitate was filtered off andrecrystallized from ethanol. Yield: 5.7 g of white crystals. The saltwas treated with 2N HCl and the free acid was extracted with ethylacetate. Yield: 3.8 g (17%).

¹H-NMR (CDCl₃): δ (ppm) 7.03-7.17 (4H, m), 5.08 (m, 1H), 4.75 (d×d, 1M),4.65 (d×d, 1H), 3.3 (d, 1H), 2.48 (d×q, 1H), 1.4-1.5 (m, 1H), 1.1-1.2(m, 1H), 0.75 (t, 3H).

EXAMPLE I3 2(S)-(4-Methoxy-phenyl)-3(R)-ethyl-pent-4-enoic acid (I3)

Step A: (4-Methoxy-phenyl)-acetic acid pent-2-enyl ester (I3a)

The ester I3a was prepared as described in Step A of example I2, usingtrans-2-penten-1-ol (49.6 ml; 487.6 mmol) and (4methoxy-phenyl)-aceticacid (81 g; 487.6 mmol) Yield: 57.7 g (52%) of a yellow oil.

¹H-NMR (CDCl₃): δ (ppm) 7.13 (d, 2H), 6.78 (d, 2H), 5.68-5.78 (m, 1H),5.42-5.52 (m, 1H), 4.45 (d, 2H), 3.72 (s, 3H), 3.5 (s, 2), 2.0 (q, 2H),0.92 (t, 3H).

Step B: 2-(4-Methoxy-phenyl)-3-ethyl-pent-4-enoic acid (I3b)

Acid I3b was prepared as described in Step B of example I2, usinghexamethyl disilazan (20.7 g, 128 mmol), n-butyl lithium (1.6M inhexane, 80 ml, 128 mmol), trimethyl chlorosilane (16.2 ml, 128 mmol),I3a (20 g, 85.4 mmol) and titanium tetrachloride (1M in dichloromethane,1.7 ml, 1.7 mmol). Yield: 15.9 g of white crystals (80%) as a racemicmixture of diastereomers (syn/anti ratio of 9/1).

Step C: 2(S)-(4-Methoxy-phenyl)-3(R)-ethyl-pent-4-enoic acid (I3)

A hot solution of the racemate I3b (15.9 g, 67.4 mmol) in ethanol (300ml) was treated with (S)-(−)-1-phenyl-ethylamine (9.4 ml, 67.4 mmol).After slowly cooling down, the precipitate was filtered off andrecrystallized from ethanol. Yield: 6.2 g of white crystals. The saltwas treated with 2N-HCl and the free acid was extracted with ethylacetate. Yield: 4.1 g (26%).

¹H-NMR (CDCl₃): δ (ppm) 7.05 (d, 2H), 6.67 (d, 2H), 5.18-5.08 (m, 1H),4.73 (d×d, 1H), 4.68 (d×d, 1H), 3.62 (s, 3H), 3.27 (d, 1H), 2.5 (m, 1H),1.4-1.5 (m, 1H), 1.1-1.2 (m, 1H), 0.75 (t, 3H).

Synthesis of the Secondary Amines VIII′

The secondary amines of the formula VIII′ were prepared by the generalMethod IB which comprises a standard coupling of an appropriate aminewith the corresponding protected carboxylic acid, followed by cleavageof the protecting group.

Method IB is illustrated by the following representative examples.

EXAMPLE I4 (S)-Pyrrolidine-2-carboxylic acid phenylamide (I4)

Step A: 2(S)-Phenylcarbamoyl-pyrrolidine-1-carboxylic acid t-butyl ester(I4a)

A solution of pyrrolidine-1,2(S)-dicarboxylic acid 1-t-butyl ester (50g, 233 mmol), EDC (45 g, 233 mmol) and HOBT (12 g, 77.5 mmol) in 1500 mlof dichloromethane was cooled to 0° C. and aniline (21 ml, 233 mmol) wasadded dropwise over a period of 15 minutes. The mixture was stirred overnight, evaporated and partitioned between ethyl acetate and 1N-HCl. Theorganic layer was washed with saturated sodium bicarbonate solution andbrine, dried over anhydrous sodium sulfate and evaporated. The crudesolid was crystallized from diethyl ether. Yield: 104 g (quantitative).

Step B: (S)-Pyrrolidine-2-carboxylic acid phenylamide (I4)

The amide I4a (64 g, 220 mmol) was treated with 150 ml of TFA (95%) at0° C. for 1 hour. The mixture was evaporated and 600 ml of 1N-HCl wasadded. After stirring for 30 minutes, the solvent was again evaporated.The crude was dried by repeated azeotropic distillation with toluene.Crystallization from diethyl ether afforded white crystals. Yield: 47 g(94%).

EXAMPLE I5 Piperidine-2(S)-carboxylic acid methylamide hydrochloride(I5)

Step A: 2(S)-Methylcarbamoyl-piperidine-1-carboxylic acid t-butyl ester(I5a)

BOC-protected L-pipecolinic acid (5 g, 21.81 mmol), which was preparedaccording to the procedure described by Ponnusamy et al. (Synthesis(1986), 48-49), was dissolved in 50 ml of THF and cooled to −75° C.Then, 1-Hydroxy-pyrrolidine-2,5-dione (2.51 g, 21.81 mmol) was added,followed by a solution of DCC (4.5 g, 21.81 mmol) in 20 ml of THF (whichwas added during a period of 45 minutes). The mixture was stirred for 3hours, then methyl amine (40% in water, 1.9 ml, 21.81 mmol) was addedand stirring continued over the weekend. The suspension was filtered,washed with ethyl acetate and evaporated. The crude residue wasredissolved in ethyl acetate and extracted with 0.1N-HCl, 5% sodiumbicarbonate and brine. After drying over anhydrous sodium sulfate andevaporation a sticky oil was obtained. Yield: 4.71 g (89%).

Step B: Piperidine-2(S)-carboxylic acid methylamide hydrochloride (I5)

Crude amide I5a (4.7 g, 19.4 mmol) was dissolved in 20 ml of dioxane andcooled in an ice-bath. Then, a 4M-solution of HCl in dioxane (9.7 ml,38.8 mmol) was added and the mixture was stirred over night. The solventwas then evaporated under reduced pressure to give a white solid. Yield:3.3 g (97%).

MS (EI): 142 [M]⁺ ¹H-NMR (DMSO-d₆): δ (ppm) 9.3 (broad s, 1H), 8.65(broad s, 1H), 8.55 (broad s, 1H), 3.7 (broad s, 1H), 3.2 (m, 1H), 2.85(m, 1H), 2.64 (d, 3H), 2.05 (m, 1H), 1.35-1.8 (m, 5H).

EXAMPLE I6 (S)-Pyrrolidine-2-carboxylic acid((S)-2-hydroxy-propyl)-amide hydrochloride (I6)

Compound I6 was prepared by EDC/HOBT-coupling of(S)-2-hydroxy-propylamine with pyrrolidine-1,2(S)-dicarboxylic acid1-t-butyl ester (analogous to Step A of Example I4), followed byHCl-cleavage in dioxane (Step B of Example I5). The crude salt was usedas such.

MS (neg. ESI): 207 [M+Cl]−, (ESI): 173 [M+H]+

EXAMPLE I7 (S)-Piperidine-2-carboxylic acid (2-methoxy-ethyl)-amidehydrochloride (I7)

Compound I7 was prepared analogous to Example I6, starting from2-methoxy-ethylamine and BOC-protected L-pipecolinic acid. The crudesalt was used as such.

MS (ESI): 187 [M+H]+

EXAMPLE I8 (S)-Piperidine-2-carboxylic acid (3-isopropoxy-propyl)-amidehydrochloride (I8)

Compound I8 was prepared analogous to Example I6, starting from3-isopropoxy-propylamine and BOC-protected L-pipecolinic acid. The crudesalt was used as such.

MS (ESI): 229 [M+H]+

EXAMPLE I9 ((S)-Piperidine-2-carboxylic acid(4-hydroxy-cyclohexyl)-amide hydrochloride (I9)

Compound I9 was prepared analogous to Example I6, starting from4-hydroxy-cyclohexylamine and BOC-protected L-pipecolinic acid.

MS (ESI): 227 [M+H]+, ¹H-NMR (DMSO-d₆): δ (ppm) 9.3 (br d, 1H), 8.62 (m,1H), 8.45 (d, 1H), 3.67 (m, 1H), 3.5 (m, 1H), 3.4 (m, 1H), 3.18 (m, 1H),2.85 (m, 1H), 2.05 (m, 1H), 1.2-1.85 (m, 14H).

EXAMPLE I10 (S)-Piperidine-2-carboxylic acid isopropylamidehydrochloride (I10)

Compound I10 was prepared analogous to Example I6, starting fromBOC-protected L-pipecolinic acid and propylamine.

¹H-NMR (DMSO-d₆): δ (ppm) 9.43 (broad s, 1H), 8.67 (broad s, 1H), 8.55(broad s, 1H), 3.86 (m, 1H), 3.67 (m, 1H), 3.19 (m, 1H), 2.85 (m, 1H),2.08 (m, 1H), 1.35-1.85 (m, 5), 1.1 (d, 3H), 1.07 (d, 3H).

EXAMPLE I11 (S)-Piperidine-2-carboxylic acid cyclopropylamidehydrochloride (I11)

Compound I11 was prepared analogous to Example I6, starting fromBOC-protected L-pipecolinic acid and cyclopropylamine.

¹H-NMR (DMSO-d₆): δ (ppm) 8.4-9.6 (broad s, 2H), 8.79 (d, 1H), 3.65 (m,1H), 3.17 (m, 1H), 2.85 (m, 1H), 2.68 (m, 1H), 2.05 (m, 1H), 1.3-1.8 (m,5H), 0.66 (m, 2H), 0.45 (m, 2H).

EXAMPLE I12 (S)-Piperidine-2-carboxylic acid benzylamide hydrochloride(I12)

Compound I12 was prepared analogous to Example I6, starting fromBOC-protected L-pipecolinic acid and benzylamine.

¹H-NMR (DMSO-d₆): δ (ppm) 9.47 (broad d, 1H), 9.22 (broad t, 1H), 8.73(broad q, 1H), 7.2-7.4 (m, 5H), 4.34 (d, 2H), 3.82 (m, 1H), 3.2 (m, 1H),2.89 (m, 1H), 2.16 (m, 1H), 1-1.85 (m, 5H).

EXAMPLE I13 (S)-Piperidine-2-carboxylic acid phenylamide hydrochloride(I13)

Compound I13 was prepared analogous to Example I6, starting fromBOC-protected L-pipecolinic acid and aniline.

¹H-NMR (DMSO-d₆): δ (ppm) 10.97 (broad s, 1H), 9.5 (broad d, 1H), 8.83(broad q, 1H), 7.68 (m, 2H), 7.35 (m, 2H), 7.1 (m, 1H), 3.98 (m, 1H),3.25 (m, 1H), 2.94 (m, 1H), 2.28 (m, 1H). 1.45-1.9 (m, 5H).

EXAMPLE I14 (S)-Piperidine-2-carboxylic acid (4-fluoro-phenyl)-amidehydrochloride (I14)

Compound I14 was prepared analogous to Example I6, starting fromBOC-protected L-pipecolinic acid and (S)-1-phenyl-ethylamine. The crudesalt was used as such.

EXAMPLE I15 (S)-3-Isobutylcarbamoyl-piperazine-1-carboxylic acid.tert.-butyl ester (I15)

Compound I15 was prepared analogous to Step A of Example I4, startingfrom isobutylamine and (S)-Piperazine-1,2,4-tricarboxylic acid 1-benzylester 4-tert-butyl ester (prepared according the procedure described inTetrahedron Letters 1989, 30, 5193) followed by heterogeneous catalytichydrogenation using Pd/C (10%) in methanol.

¹H-NMR (DMSO-d₆): δ (ppm) 7.83 (broad s, 1H), 2.7-4.2 (very broadmultiplets, 9H), 1.7 (m, 1H), 1.39 (s, 9H), 0.82 (d, 3H).

Synthesis of the Hydroxamic Acids I′″

The hydroxamic acids of the formula I′″ were prepared by the generalMethod A which comprises the coupling of intermediates VII′ with thecorresponding amines VIII′ or their appropriately protected forms,followed by an ozonolysis of the vinyl olefin, coupling withO-(trimethylsilyl)-hydroxylamine and deprotection if required.

Method A is illustrated by the following representative example 1.

EXAMPLE 13(S)-(4-Chloro-phenyl)-2(S)-ethyl-N-hydroxy-4-morpholin-4-yl-4-oxo-butyramide(1)

Step A:2(S)-(4-Chloro-phenyl)-3(S)-ethyl-1-morpholin-4-yl-pent-4-en-1-one (1a)

Morpholine (0.107 ml, 1.23 mmol) and the intermediate I2 (239 mg, 1.12mmol) were dissolved in 5 ml of DMF and cooled in an ice-bath. HOBT (188mg, 1.23 mmol), triethylamine (0.309 ml, 2.24 mmol) and EDC (214 mg,1.12 mmol) were added successively. The ice-bath was removed andstirring continued over night. The mixture was evaporated, redissolvedin diethyl ether and extracted with 2N-HCl (twice), 5% sodiumbicarbonate and brine. The organic layer was dried over anhydrous sodiumsulfate and evaporated to give a pale yellow oil. Yield (crude): 320 mg(93%).

MS (ESI): 308.1 [M+H]⁺ ¹H-NMR (CDCl₃): δ (ppm) 7.05-7.2 (m, 4H),5.12-5.24 (m, 1H), 4.6-4.77 (m, 2H), 3.1-3.65 (m, 9H), 2.69 (q×d, 1H),1.54 (m, 1H), 1.12 (m, 1H), 0.8 (t, 3H).

Step B: 3(S)-(4-Chloro-phenyl)-2(S)-ethyl-4-morpholin-4-yl-4-oxo-butyricacid (1b)

The crude olefin 1a (315 mg, 1.02 mmol) was dissolved in 15 ml ofmethanol and cooled to −75° C. A constant flow (50 l/h) of ozone wasintroduced at a temperature around −70° C. After about 10 minutes allstarting material was consumed a blue colour appeared. The solution wasthen purged with nitrogen and dimethylsulfide (0.375 ml, 5.1 mmol) wasadded. The cooling bath was removed and stirring continued for 2 hours.The solvents were evaporated and the crude oil redissolved in 7 ml oft-butanol. A solution of sodium dihydrogenphosphate monohydrate (423 mg,3.06 mmol) in 2 ml of water was added, followed by 2-methyl-2-butene(0.54 ml, 5.1 mmol) and sodium chlorite (185 mg, 2.04 mmol) in 2 ml ofwater. The mixture turns yellow and an exothermic reaction is observed(eventual cooling with ice-bath may be required for large scalereactions). After 1 hour the reaction had ceased and the colourlesssolution was treated with sodium sulfite (129 mg, 1.02 mmol) in 2 ml ofwater. Most of the solvents were then evaporated and the residue waspartitioned between diethyl ether and 1N-HCl. The organic layer wasseparated and extracted twice with 1N-NaOH. The aqueous layers werecombined, acidified with cold concentrated HCl and extracted three timeswith diethyl ether. The combined organic layers were then washed withwater and brine and dried over anhydrous sodium sulfate. Afterevaporation a white foam was obtained. Yield (crude): 318 mg (95 %).

MS (ESI): 326.1 [M+H]⁺ ¹H-NMR (CDCl₃): δ (ppm) 7.1-7.3 (m, 4H), 4.82 (d,1H), 3.2-3.7 (m, 8H), 3.07 (d×d×d, 1H), 1.6 (m, 2H), 0.88 (t, 3H).

Step C:3(S)-(4-Chloro-phenyl)-2(S)-ethyl-N-hydroxy-4-morpholin-4-yl-4-oxo-butyramide(1)

The crude acid 1b (310 mg, 0.95 mmol) was dissolved in 5 ml ofdichloromethane. HOPO (111 mg, 1 mmol) and MEI (0.159 ml, 1.14 mmol)were added successively. The mixture was stirred for 30 minutes, thenTMSONH2 (0.233 ml, 1.9 mmol) was added. After one hour another portionof TMSONH2 (0.233 ml, 1.9 mmol) was added and stirring continued overnight. The reaction mixture was filtered (solid contained mostly HOPO)diluted with 20 ml of dichloromethane and extracted twice with 2N-HCland brine, dried over anhydrous sodium sulfate and evaporated. The crudesolid (237 mg, 73%) was purified by preparative HPLC(methanol/water=1/1). Yield: 100 mg (31%).

MS (ESI): 341.1 [M+H]⁺ ¹H-NMR (DMSO-d₆): δ (ppm) 10.28 (s, 1H), 8.55 (s,1H), 7.32 (s, 4H), 4.12 (d, 1H), 3.15-3.75 (m, 8H), 2.77 (t×d, 1H), 1.45(m, 2H), 0.82 (t, 3H).

EXAMPLE 22(R)-Benzyloxymethyl-4-[4-(4-chloro-phenyl)-piperazin-1-yl]-N-hydroxy-3(S)-(4-methoxy-phenyl)-4-oxo-butyramide(2)

Compound 2 was prepared analogous to steps A to C of example 1, startingfrom 1-(4-chloro-phenyl)-piperazine and intermediate I1.

MS (neg. ESI): 536.2 [M−H]⁻ ¹H-NMR (DMSO-d₆): δ (ppm) 10.3 (s, 1H), 8.57(s, 1H), 7.15-7.37 (m, 9H), 6.87 (d, 2H), 6.82 (d, 2H), 4.45 (AB system,2H), 4.11 (d, 1H), 3.77 (m, 1H), 3.72 (s, 3H), 3.38-3.66 (m, 5H), 3.26(m, 1H), 2.95 (m, 3H), 2.68 (m, 1H).

EXAMPLE 32(R)-Benzyloxymethyl-N-hydroxy-3(S)-(4-methoxy-phenyl)-4-oxo-4-piperidin-1-yl-butyramide(3)

Compound 3 was prepared analogous to steps A to C of example 1, startingfrom piperidine and intermediate I1.

MS (ESI): 427.2 [M+H]⁺, 449.2 [M+Na]⁺ ¹H-NM (DMSO-d₆): δ (ppm) 10.29 (s,1H), 8.56 (s, 1H), 7.25-7.4 (m, 5H), 7.21 (d, 2H), 6.81 (d, 2H), 4.45(AB system, 2H), 3.72 (s, 3H), 3.3-3.65 (m, 6H), 3.23 (m, 1H), 0.85-1.55(m, 6H).

EXAMPLE 4N-Hydroxy-2(R)-hydroxymethyl-3(S)-(4-methoxy-phenyl)-4-oxo-4-piperidin-1-yl-butyramide(4)

A solution of hydroxamic acid 3 (330 mg, 0.64 mmol) in 20 ml of methanolwas hydrogenated under normal pressure with 70 mg of palladium on bariumsulfate for about 5 hours. After filtration of the catalyst the solventwas evaporated and the crude product was purified by preparative HPLC togive a white solid. Yield: 133 mg (62%).

MS (ESI): 337.0 [M+H]⁺, 359.0 [M+Na]⁺ ¹H-NMR (DMSO-d₆): δ (ppm) 10.2(broad s, 1H), 8.48 (broad s, 1H), 7.2 (d, 2H), 6.8 (d, 2H), 4.65 (broads, 1H), 4.0 (d, 1H), 3.3-3.6 (m, 6H), 2.97 (m, 1H), 0.9-1.7 (m, 6H).

EXAMPLE 5(S)-4-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-3-isobutylcarbamoyl-piperazine-1-carboxylicacid .tert-butyl ester (5)

Compound 5 was prepared analogous to Steps A to C of Example 1, startingfrom amine I15 and succinate I2.

MS (neg. ESI): 537.0 [M−H]⁻ MS (pos. ESI): 561.0 [M+Na]⁺, 577.0 [M+K]⁺,461.0 [M−BOC]⁺ ¹H-NMR (DMSO-d₆): 2 rotamers at 20° C. (ratio 3:2) δ(ppm) 10.25 (broad s, 1H), 8.56 and 8.53 (broad s, 1H), 8.27 and 7.66(broad s, 1H), 7.2-7.4 (m, 4H), 4.73 and 4.53 (broad d, 1H), 2.4-4.25(overlapping broad multiplets, 10H), 1.2-1.85 (overlapping broadmultiplets, 3H), 1.35 (s, 9H), 0.7-1.0 (overlapping multiplets, 9H).

EXAMPLE 6(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperazine-2-carboxylicacid isobutyl-amide trifluoro-acetate (6)

A solution of 5 (20 mg, 0.037 mmol) in 2.5 ml of dichloromethane wascooled in an ice-bath. It was then treated with TFA (29 μl, 0.37 mmol)and stirred at room temperature over night. After addition of 5 ml oftoluene, the mixture was evaporated to give the crude TFA-salt.

Yield: 18 mg (88%). MS (neg. ESI): 437.0 [M−H]⁻, 551.0 [M+CF₃COO]⁻ MS(pos. ESI): 439.0 [M+H]⁺, 406.0 [M−NH−OH]⁺ ¹H-NMR (DMSO-d₆): 2 rotamersat 20° C. (ratio 3:1) δ (ppm) 10.3 and 10.28 (broad s, 1H), 9.24 and 9.0and 8.57 and 8.23 (very broad singlets, together 3H), 8.41 and ˜7.3overlapping with aromatic signals (broad t, 1H), 7.15-7.45 (m, 4H), 5.11and 4.92 (broad d, 1H), 4.4 and 4.37 (broad d, 1H), 4.23 and 4.01 (d,1H), 2.6-3.3 (overlapping multiplets, 8H), 1.5 (m, 2H), 0.55-1.25(overlapping multiplets, 9H).

EXAMPLE 71-[4-Benzyloxy-3(R)-hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-butyryl]-piperidine-2(S)-carboxylicacid methylamide (7)

Compound 7 was prepared analogous to steps A to C of example 1, startingfrom piperidine-2(S)-carboxylic acid methylamide hydrochloride, I5, andintermediate I1.

MS (ESI): 484.2 [M+H]⁺, 506.2 [M+Na]⁺ ¹H-NMR (DMSO-d₆): 2 rotamers at20° C. (ratio 2:3) δ (ppm) 10.3 and 10.26 (s, 1H), 8.56 and 8.53 (s,1H), 7.83 and 7.13 (broad q, 1H), 7.25-7.6 (m, 5H), 7.21 (m, 2H), 6.83(m, 2H), 4.93 and 4.6 (m, 1H), 4.36-4.56 (m, 2H), 4.32 and 3.96 (m, 1H),4.1 and 3.79 (d, 1H), 3.73 (s, 3H), 2.96-3.68 (m, 4H), 2.62 and 2.47 (d,3H), 1.94 (m, 1H), 0.65-1.65 (m, 5H).

EXAMPLE 81-[4-Hydroxy-3(R)-hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-butyryl]-piperidine-2(S)-carboxylicacid methylamide (8)

A solution of hydroxamic acid 7 (100 mg, 0.21 mmol) in 20 ml of methanolwas hydrogenated under normal pressure with 50 mg of palladium on bariumsulfate for about 5 hours. After filtration of the catalyst the solventwas evaporated and the crude product was purified by preparative HPLC togive a white solid. Yield: 53 mg (65%).

MS (ESI): 394.2 [M+H]⁺, 416.1 [M+Na]⁺ ¹H-NMR (DMSO-d₆): 2 rotamers at20° C. (ratio 3:2) δ (ppm) 10.18 and 10.16 (s, 1H), 8.46 and 8.44 (s,1H), 7.4 and 7.13 (broad q, 1H), 7.2 (m, 2H), 6.8 (m, 2H), 4.97 and 4.74(m, 1H), 4.66 (broad s, 1H), 4.33 and 3.99 (m, 1H), 4.09 and 3.84 (d,1H), 3.72 (s, 3H), 3.4-3.6 (m, 2H), 2.9-3.15 and 2.55-2.7 (m, 2H), 2.67and 2.48 (d, 3H), 1.98 (m, 1H), 0.75-1.65 (m, 5M).

EXAMPLE 91-[3(S)-Hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-pentanoyl]-piperidine-2S-carboxylicacid methylamide (9)

Compound 9 was prepared analogous to steps A to C of example 1, startingfrom piperidine-2(S)-carboxylic acid methylamide hydrochloride amine I5,and intermediate I3.

MS (neg. ESI): 390.1 [M−H]⁻, 425.8 [M−Cl]⁻ ¹H-NMR (DMSO-d₆): 2 rotamersat 20° C. (ratio 1:1) δ (ppm) 10.23 and 10.18 (s, 1H), 7.91 and 7.15(broad q, 1H), 7.19 (m, 2H), 6.8 (m, 2H), 4.98 and 4.62 (m, 1H), 4.33and 4.03 (m, 1H), 4.07 and 3.77 (d, 1H), 3.72 (s, 3H), 2.6-3.2 (m, 2H),2.66 and 2.47 (d, 3H), 2.01 and 1.91 (d, 1H), 1.1-1.7 (m, 8H), 0.7-0.90(m, 3H).

EXAMPLE 10(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid cyclopropylamide (10)

Compound 10 was prepared analogous to steps A to C of example 1,starting from amine I11 and succinate I3.

MS (neg. ESI): 416.1 [M−H]⁻ MS (pos. ESI): 440.1 [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 1:1) δ (ppm) 10.23 and 10.2 (s,1H), 8.53 and 8.47 (s, 1H), 8.07 and 7.08 (broad d, 1H), 7.2 (m, 2H),6.7 (m, 2H), 4.93 and 4.6 (m, 1H), 3.9-4.4 (overlapping multiplets, 3H),3.74 and 3.72 (s, 3H), 2.65-3.15 (overlapping multiplets, 2H), 1.98 and1.88 (broad d, 1H), 0.1-1.7 (overlapping multiplets, 14H).

EXAMPLE 11(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid (2-methoxy-ethyl)-amide (11)

Compound 11 was prepared analogous to steps A to C of example 1,starting from amine I7 and succinate I3.

MS (neg. ESI): 390.1 [M−H]⁻, 425.8 [M−Cl]⁻ ¹H-NMR DMSO-d₆): 2 rotamersat 20° C. (ratio 1:1) δ (ppm) 10.23 and 10.18 (s, 1H), 7.91 and 7.15(broad q, 1H), 7.19 (m, 2H), 6.8 (m, 2H), 4.98 and 4.62 (m, 1H), 4.33and 4.03 (m, 1H), 4.07 and 3.77 (d, 1H), 3.72 (s, 3H), 2.6-3.2 (m, 2H),2.66 and 2.47 (d, 3H), 2.01 and 1.91 (d, 1H), 1.1-1.7 (m, 8H), 0.7-0.90(m, 3H).

EXAMPLE 12(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid (4-hydroxy-cyclohexyl)-amide (12)

Compound 12 was prepared analogous to steps A to C of example 1,starting from amine I9 and succinate I3.

MS (neg. ESI): 390.1 [M−H]⁻, 425.8 [M−Cl]⁻ ¹H-NMR (DMSO-d₆): 2 rotamersat 20° C. (ratio 1:1) δ (ppm) 10.23 and 10.18 (s, 1H), 7.91 and 7.15(broad q, 1H), 7.19 (m, 2H), 6.8 (m, 2H), 4.98 and 4.62 (m, 1H), 4.33and 4.03 (m, 1H), 4.07 and 3.77 (d, 1H), 3.72 (s, 3H), 2.6-3.2 (m, 2H),2.66 and 2.47 (d, 3H), 2.01 and 1.91 (d, 1H), 1.1-1.7 (m, 8H), 0.7-0.90(m, 3H).

EXAMPLE 13(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid benzylamide (13)

Compound 13 was prepared analogous to steps A to C of example 1,starting from amine I12 and succinate I3.

MS (neg. ESI): 466.1 [M−H]⁻ MS (pos. ESI): 490.0 [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 1:1) δ (ppm) 10.22 and 10.19 (s,1H), 8.52 and 8.48 (s, 1H), 8.59 and 7.7 (broad t, 1H), 6.7-7.4(overlapping multiplets, 9H), 5.06 and 4.76 (m, 1H), 3.7-4.4(overlapping multiplets, 4H), 3.73 and 3.70 (s, 3H), 3.05 and 2.92 (m,1H), 2.72 (m, 1H), 2.08 and 1.97 (broad d, 1H), 1.2-1.7 (m, 6H), 0.95(broad m, 1H), 0.82 and 0.76 (t, 3H).

EXAMPLE 14(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid (4-fluoro-phenyl)-amide (14)

Compound 14 was prepared analogous to steps A to C of example 1,starting from amine I14 and succinate I3.

MS (neg. ESI): 470.0 [M−H]⁻ MS (pos. ESI): 494.1 [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 2:1) δ (ppm) 10.21 (broad s, 1H),10.06 and 9.73 (s, 1H), 8.51 and 8.47 (broad s, 1H), 6.7-7.7(overlapping multiplets, 8H), 5.08 and 4.88 (m, 1H), 4.35 and 4.07(broad d, 1H), 4.127 and 3.84 (d, 1H), 3.74 and 3.71 (s, 3H), 2.0-3.6(overlapping multiplets, 3H), 0.95-1.75 (overlapping multiplets, 7H),0.7-0.95 (overlapping multiplets, 3H).

EXAMPLE 15(S)-1-[(2S,3S)-2-(4-Chloro-phenyl-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid isopropylamide (15)

Compound 15 was prepared analogous to steps A to C of example 1,starting from amine I10 and succinate I2.

MS (neg. ESI): 422.0 [M−H]⁻ MS (pos. ESI): 446.0 [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 3:2) δ (ppm) 10.28 and 10.25 (s,1H), 8.55 and 8.50 (s, 1H), 7.84 and 6.77 (d, 1H), 7.25-7.5 (m, 4H),4.95 and 4.65 (broad d, 1H), 4.3 and 4.08 (broad d, 1H), 4.17 and 3.89(d, 1H), 3.92 and 3.72 (m, 1H), 3.0 (m, 1H), 2.75 (m, 1H), 2.05 and 1.93(broad d, 1H), 1.2-1.7 (overlapping multiplets, 7H), 0.7-1.2(overlapping multiplets, 9H).

EXAMPLE 16(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid cyclopropylamide (16)

Compound 16 was prepared analogous to steps A to C of example 1,starting from amine I11 and succinate I2.

MS (neg. ESI): 420.1 [M−H]⁻ MS (pos. ESI): 44.0 [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 1:1) δ (ppm) 10.25 (broad s, 1H),8.55 and 8.51 (broad s, 1H), 8.09 and 7.48 (broad d, 1H), 7.3 (m, 4H),4.91 and 4.6 (broad s, 1H), 4.28 and 4.03 (broad d, 1H), 4.15 and 3.84(d, 1H), 3.08 and 2.94 (m, 1H), 2.7 (m, 1H), 1.97 and 1.89 (broad d,1H), 0.15-1.75 (overlapping multiplets, 15H).

EXAMPLE 17(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid (3-isopropoxy-propyl)-amide (17)

Compound 17 was prepared analogous to steps A to C of example 1,starting from amine I8 and succinate I2.1

MS (neg. ESI): 390.1 [M−M]⁻, 425.8 [M−Cl]⁻ ¹H-NMR (DMSO-d₆): 2 rotamersat 20° C. (ratio 1:1) δ (ppm) 10.23 and 10.18 (s, 1H), 7.91 and 7.15(broad q, 1H), 7.19 (m, 2H), 6.8 (m, 2H), 4.98 and 4.62 (m, 1H), 4.33and 4.03 (m, 1H), 4.07 and 3.77 (d, 1H), 3.72 (s, 3H), 2.6-3.2 (m, 2H),2.66 and 2.47 (d, 3H), 2.01 and 1.91 (d, 1H), 1.1-1.7 (m, 8H), 0.7-0.90(m, 3H).

EXAMPLE 18(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid (4-hydroxy-cyclohexyl)-amide (18)

Compound 18 was prepared analogous to steps A to C of example 1,starting from amine I9 and succinate I2.

MS (neg. ESI): 390.1 [M−H]⁻, 425.8 [M−Cl]⁻ ¹H-NMR (DMSO-d₆): 2 rotamersat 20° C. (ratio 1:1) δ (ppm) 10.23 and 10.18 (s, 1H), 7.91 and 7.15(broad q, 1H), 7.19 (m, 2H), 6.8 (m, 2H), 4.98 and 4.62 (m, 1H), 4.33and 4.03 (m, 1H), 4.07 and 3.77 (d, 1H), 3.72 (s, 3H), 2.6-3.2 (m, 2H),2.66 and 2.47 (d, 3H), 2.01 and 1.91 (d, 1H), 1.1-1.7 (m, 8H), 0.7-0.90(m, 3H).

EXAMPLE 19(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-2-carboxylicacid benzylamide (19)

Compound 19 was prepared analogous to steps A to C of example 1,starting from amine I12 And succinate I2

MS (neg. ESI): 470.0 [M−H]⁻ MS (pos. ESI): 494.0 [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 55:45) δ (ppm) 10.25 (broad s,1H), 8.55 and 8.52 (broad s, 1H), 8.6 and 7.95 (broad t, 1H), 7.05-7.65(m, 9H), 5.04 and 4.78 (broad d, 1H), 3.88-4.42 (overlapping multiplets,4H), 3.08 and 2.95 (m, 1H), 2.74 (m, 1H) 2.07 and 1.98 (broad d, 1H),1.21.7 (m, 6H), 0.97 (m, 1H), 0.72 (t, 3H).

EXAMPLE 20(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid phenylamide (20)

Compound 20 was prepared analogous to steps A to C of example 1,starting from amine I13 And succinate I2.

MS (neg. ESI): 446.0 [M-H]⁻ MS (pos. ESI): 480. [M+Na]⁺ ¹H-NMR(DMSO-d₆): 2 rotamers at 20° C. (ratio 2:1) δ (ppm) 10.25 (broad s, 1H),10.05 and 9.7 (broad s, 1H), 8.5 (broad s, 1H), 6.95-7.7 (m, 9H), 5.08and 4.9 (broad d, 1H), 4.34 and 4.07 (broad d, 1H), 4.17 and 3.93 (d,1H), 3.23 and 3.09 (m, 1H), 2.72 (m, 1H), 0.90-2.15 (overlappingmultiplets, 8H), 0.82 and 0.74 (t, 3H).

EXAMPLE 211-[3(S)-Hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-pentanoyl]-pyrrolidine-2(S)-carboxylicacid phenylamide (21)

Compound 21 was prepared analogous to steps A to C of example 1,starting from amine I4 and succinate I3.

MS (neg. ESI): 438.0 [M−H]⁻, 474.0 [M−Cl]⁻ ¹H-NMR DMSO-d₆): 2 rotamersat 20° C. (ratio 2:1) δ (ppm) 10.3 and 10.22 (s, 1H), 10.15 and 9.86 (s,1H), 8.5 and 8.42 (s, 1H), 6.7-7.7 (m, 9H), 4.47 (m, 1H), 3.85 and 3.5(d, 1H), 3.68-3.78 and 3.57 (m, 1H), 3.72 (s, 3H), 3.46 and 3.28 (m,1H), 2.56-2.75 (m, 1H), 1.2-2.65 (m, 6H), 0.81 and 0.74 (t, 3H).

EXAMPLE 22(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-pyrrolidine-2-carboxylicacid ((S)-2-hydroxy-propyl)-amide (22)

Compound 22 was prepared analogous to steps A to C of example 1,starting from amine I6and intermediate I2.

MS (neg. ESI): 442.2 [M−H]⁻ ¹H-NMR (DMSO-d₆): 2 rotamers at 20° C.(ratio 2:1) δ (ppm) 10.34 and 10.26 (s, 1H), 10.2 and 9.9 (s, 1H), 8.55and 8.45 (s, 1H), 6.95-7.7 (m, 9H), 4.46 (m, 1H), 3.93 and 3.56 (d, 1H),3.74 and 3.59 (m, 1H), 3.44 and 3.28 (m, 1H), 2.58-2.77 (m, 1I),1.2-2.45 (m, 6H), 0.82 and 0.75 (t, 3H).

The table below represents the above examples with the general formulaIa

I_(a)

Exp. n Zi Ai Ri₁ Xi Ri₃ 1 1 O H Ethyl Cl 2 1 N-4-Chlorophenyl HCH₂—O-Benzyl OMe 3 1 CH₂ H CH₂—O-Benzyl OMe 4 1 CH₂ H CH₂—OH OMe 5 1N—BOC —CO—NH— Ethyl Cl iso-Butyl 6 1 NH —CO—NH— Ethyl Cl iso-Butyl 7 1CH₂ —CO—NH— CH₂—O-Benzyl OMe Methyl 8 1 CH₂ —CO—NH— CH₂—OH OMe Methyl 91 CH₂ —CO—NH— Ethyl OMe Methyl 10 1 CH₂ —CO—NH— Ethyl OMe Cyclo-Propyl11 1 CH₂ —CO—NH— Ethyl OMe 2-Methoxy-ethyl 12 1 CH₂ —CO—NH— Ethyl OMe4-Hydroxy-cyclohexyl 13 1 CH₂ —CO—NH— Ethyl OMe Benzyl 14 1 CH₂ —CO—NH—Ethyl OMe 4-Fluoro-phenyl 15 1 CH₂ —CO—NH— Ethyl Cl iso-Propyl 16 1 CH₂—CO—NH— Ethyl Cl cyclo-Propyl 17 1 CH₂ —CO—NH— Ethyl Cl3-Isopropoxy-propyl 18 1 CH₂ —CO—NH— Ethyl Cl 4-Hydroxy-cyclohexyl 19 1CH₂ —CO—NH— Ethyl Cl Benzyl 20 1 CH₂ —CO—NH— Ethyl Cl Phenyl 21 0 CH₂—CO—NH— Ethyl OMe Phenyl 22 0 CH₂ —CO—NH— Ethyl Cl (S)-2-Hydroxy-propyl

1. A compound of Formula I

wherein R₁ is lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl orC₄-C₁₈aryl each of which is independently optionally substituted byhydroxy, halogen, lower alkoxy, C₃-C₈cycloalkyl-lower alkoxy, or C₄-C₁₈aryl-lower alkoxy; X is halogen, cyano, lower alkyl, halo-substitutedlower alkyl, C₄-C₁₈aryl, C₄-C₁₈aryl-lower alkyl, hydroxy, —OR₅, SR₅ or—NR₆R₇, each of which is optionally substituted by halogen, hydroxy,lower alkoxy, C₃-C₆cycloalkyl-lower alkoxy, or C₄-C₁₈aryl-lower alkoxywherein R₅ is hydrogen, lower alkyl, C₃-C₈cycloalkyl,C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl and R₆ and R₇ are independently H,lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl; Z is—CH₂—, —CHR₈—, —O—, —S—, or —N(R₈)— wherein R₈ is H, lower alkyl,C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl, C₄-C₁₈aryl lower alkoxycarbonylor C₄-C₈aryloxycarbonyl, each of which is independently optionallysubstituted by halogen, hydroxy, lower alkoxy, C₃-C₆cycloalkyl-loweralkoxy, or C₄-C₈aryl-lower alkoxy; A is hydrogen, —CR₁₀R₁₁-Q-R₁₂,—C(O)-Q-R₁₂ or —C(S)-Q-R₁₂ wherein R₁₀ and R₁₁ are independently H,lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl eachof which is independently optionally substituted by halogen, hydroxy,lower alkoxy, C₃-C₆cycloalkyl-lower alkoxy, or C₄-C₁₈ aryl-lower alkoxy,Q is —NR₈—, —S— or —O—, where R₈ is as defined above, and R₁₂ is loweralkyl C₃-C₈cycloalkyl, C₄-C₁₈aryl, C₄-C₁₈aryl-lower alkyl, eachoptionally substituted by hydroxy, halogen, lower alkoxy,C₃-C₆cycloalkyl, C₃-C₆cycloalkoxy, C₄-C₁₈aryl or C₄-C₁₈aryl-loweralkoxy; and R₃ and R₄ is Hydrogen or lower alkyl; and n is 0 or 1, or apharmaceutically-acceptable and -cleavable ester thereof or acidaddition salts thereof.
 2. A compound according to claim 1 of formula II

wherein R₁′ is H, lower alkyl or C₃-C₈cycloalkyl, each of which isoptionally substituted by hydroxy, halogen, lower alkoxy orC₄-C₁₈aryl-lower alkoxy; X′ is halogen, cyano, lower alkyl,halo-substituted lower alkyl or lower alkoxy, each of which isoptionally substituted by halogen, hydroxy or lower alkoxy; Z′ is —CH₂—or —N(R′₈)— wherein R′₈ is H, lower alkyl, C₄-C₁₈aryl (optionallysubstituted by halogen), lower alkoxycarbonyl or C₄-C₁₈aryloxycarbonyl;A′ is H or —C(O)-Q′-R₁₂′ wherein Q′ is —S— or —O— and R₁₂′ is loweralkyl, C₃-C₈ cycloalkyl, C₄-C₁₈aryl, each optionally substituted byhydroxy, halogen, lower alkoxy, C₃-C₈cycloalkyl, or C₄-C₁₈aryl, or apharmaceutically acceptable and cleavable ester thereof or acid additionsalts thereof.
 3. A compound according to claim 1 of formula I′ orformula II′

wherein the symbols are as defined above.
 4. A compound according toclaim 1 selected from:3(S)-(4-Chloro-phenyl)-2(S)-ethyl-N-hydroxy-4-morpholin-4-yl-4-oxo-butyramide;2(R)-Benzyloxymethyl-4-[4-(4-chloro-phenyl)-piperazin-1-yl]-N-hydroxy-3(S)-(4-methoxy-phenyl)-4-oxo-butyramide;2(R)-Benzyloxymethyl-N-hydroxy-3(S)-(4-methoxy-phenyl)-4-oxo-4-piperidin-1-yl-butyramide,N-Hydroxy-2(R)-hydroxymethyl-3(S)-(4-methoxy-phenyl)-4-oxo-4-piperidin-1-yl-butyramide;(S)-4-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-3-isobutylcarbamoyl-piperazine-1-carboxylicacid tert-butyl ester;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperazine-2-carboxylicacid isobutyl-amide trifluoro-acetate;1-[4-Benzyloxy-3(R)-hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-butyryl]-piperidine-2(S)-carboxylicacid methylamide;1-[4-Hydroxy-3(R)-hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-butyryl]-piperidine-2(S)-carboxylicacid methylamide;1-[3(S)-Hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-pentanoyl]-piperidine-2(S)-carboxylicacid methylamide;(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid cyclopropylamide;(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid (2-methoxy-ethyl)-amide;(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid (4-hydroxy-cyclohexyl)-amide;(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid benzylamide;(S)-1-[(2S,3S)-3-Hydroxycarbamoyl-2-(4-methoxy-phenyl)-pentanoyl]-piperidine-2-carboxylicacid (4-fluoro-phenyl)-amide;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid isopropylamide;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid cyclopropylamide;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid (3-isopropoxy-propyl)-amide;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid (4-hydroxy-cyclohexyl)-amide;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid benzylamide;(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-piperidine-2-carboxylicacid phenylamide;1-[3(S)-Hydroxycarbamoyl-2(S)-(4-methoxy-phenyl)-pentanoyl]-pyrrolidine-2(S)-carboxylicacid phenylamide; and(S)-1-[(2S,3S)-2-(4-Chloro-phenyl)-3-hydroxycarbamoyl-pentanoyl]-pyrrolidine-2-carboxylicacid ((S)-2-hydroxy-propyl)-amide; or a pharmaceutically acceptable andcleavable ester thereof of acid addition salts thereof.
 5. A method ofinhibiting production of soluble TNF in a subject in need of suchtreatment which method comprises administering to said subject aneffective amount of a compound according to claim
 1. 6. A pharmaceuticalcomposition comprising a compound according to claim 1 in associationwith a pharmaceutically acceptable diluent or carrier.
 7. A method ofinhibiting neuropathic pain in a subject in need of such treatment whichmethod comprises administering to said subject an effective amount of acompound according to claim
 1. 8. A process for the preparation of acompound of formula I

wherein R₁ is lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl orC₄-C₁₈aryl each of which is independently optionally substituted byhydroxy, halogen, lower alkoxy, C₃-C₈cycloalkyl-lower alkoxy, or C₄-C₁₈aryl-lower alkoxy; X is halogen, cyano, lower alkyl, halo-substitutedlower alkyl, C₄-C₁₈aryl, C₄-C₁₈aryl-lower alkyl, hydroxy, —OR₅, SR₅ or—NR₆R₇, each of which is optionally substituted by halogen, hydroxy,lower alkoxy, C₃-C₆cycloalkyl-lower alkoxy, or C₄-C₁₈aryl-lower alkoxywherein R₅ is hydrogen, lower alkyl, C₃-C₈cycloalkyl,C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl and R₆ and R₇ are independently H,lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl;C₁₈heterocycloalkyl or C₄-C₁₈aryl; Z is —CH₂—, —CHR₈—, —O—, —S—, or—N(R₈)— wherein R₈ is H, lower alkyl, C₃-C₈cycloalkyl,C₃-C₁₈heterocycloalkyl, C₄-C₁₈aryl lower alkoxycarbonyl orC₄-C₈aryloxycarbonyl, each of which is independently optionallysubstituted by halogen, hydroxy, lower alkoxy, C₃-C₆cycloalkyl-loweralkoxy, or C₄-C₈aryl-lower alkoxy; A is hydrogen, —CR₁₀R₁₁-Q-R₁₂,—C(O)-Q-R₁₂ or —C(S)-Q-R₁₂ wherein R₁₀ and R₁₁ are independently H,lower alkyl, C₃-C₈cycloalkyl, C₃-C₁₈heterocycloalkyl or C₄-C₁₈aryl eachof which is independently optionally substituted by halogen, hydroxy,lower alkoxy, C₃-C₆cycloalkyl-lower alkoxy, or C₄-C₁₈ aryl-lower alkoxy,Q is —NR₈—, —S— or —O—, where R₈ is as defined above, and R₁₂ is loweralkyl C₃-C₈cycloalkyl, C₄-C₁₈aryl, C₄-C₁₈aryl-lower alkyl, eachoptionally substituted by hydroxy, halogen, lower alkoxy,C₃-C₆cycloalkyl, C₃-C₆cycloalkoxy, C₄-C₁₈aryl or C₄-C₁₈aryl-loweralkoxy; and R₃ and R₄ is Hydrogen or lower alkyl; and n is 0 or 1, or apharmaceutically-acceptable and -cleavable ester thereof or acidaddition salts thereof which process comprises converting acorresponding free carboxylic acid derivative of formula V

wherein the symbols are as for Formula I, to the correspondinghydroxamic acid derivative of formula I.
 9. A method of inhibitingmatrix metalloproteinase activity in a subject in need of such treatmentwhich method comprises administering to said subject an effective amountof a compound according to claim
 1. 10. A method of reducinginflammation in a subject in need of such treatment which methodcomprises administering to said subject an effective amount of acompound according to claim
 1. 11. A method of inducingimmunosuppression in a subject in need of such treatment which methodcomprises administering to said subject an effective amount of acompound according to claim
 1. 12. A method of treating neuropathic painin a subject in need of such treatment which method comprisesadministering to said subject an effective amount of a compoundaccording to claim 1.